Mechanisms and implications of intracellular Zn2+ release in acidified neurons

描述（由申请人提供）：升高的细胞内游离Zn 2+浓度（[Zn 2 +]i）与中风引起的神经元死亡有关。因此，恢复低[Zn2+]i的疗法有可能改善中风的结局。拟议研究的目标是设计这样一种疗法。对海马神经元的初步研究表明，类似于中风期间发生的细胞内酸化导致[Zn 2]i升高，这是由于Zn 2+从细胞内储存中释放。我们将确定低亲和力Zn2+配体，如腺嘌呤核苷酸，在这些[Zn2+]i的海拔，并建立是否可以通过酸诱导的[Zn2+]i海拔的特异性螯合改善神经元的生存能力。特别是，我们将设计一种专门针对这些酸诱导的[Zn2+]i升高的Zn2+螯合方法。这项研究将在小鼠的原代海马神经元上进行。目的1将阐明当pH下降时，哪些低亲和力的细胞内Zn 2+配体释放该离子 进入中风特征范围。在目标2中，将开发一种方法来特异性螯合酸诱导的[Zn2+]i升高。目的3将确定这种锌离子螯合方法是否能提高缺氧缺糖神经元的活力。如果是这样的话，我们的方法将提供一个框架，为发展的治疗，抵消过度[Zn2+]i海拔在中风。

DESCRIPTION (provided by applicant): Elevated intracellular free Zn2+ concentrations ([Zn2+]i) have been implicated in the neuronal death caused by stroke. Therefore, a therapy that restores low [Zn2+]i has the potential to improve the outcome of stroke. The goal of proposed research is to design such a therapy. Preliminary studies on hippocampal neurons indicate that an intracellular acidification similar to that taking place during stroke causes [Zn2]i elevations due to Zn2+ release from intracellular stores. We will determine the role of low affinit Zn2+ ligands, such as adenine nucleotides, in these [Zn2+]i elevations and establish whether neuronal viability can be improved by a specific chelation of acid-induced [Zn2+]i elevations. In particular, we will design a method of Zn2+ chelation that specifically targets these acid-induced [Zn2+]i elevations. The research will be conducted on primary hippocampal neurons from mice. Aim 1 will clarify which low affinity intracellular Zn2+ ligands release this ion when the pH drops into a stroke-characteristic range. In Aim 2, a method will be developed to specifically chelate acid-induced [Zn2+]i elevations. Aim 3 will determine whether this method of Zn2+ chelation improves the viability of neurons exposed to oxygen and glucose-deprivation. If it does, our method will provide a framework for the development of therapy that counteracts excessive [Zn2+]i elevations during stroke.